Example 1
Adult fish were raised and maintained as described in [28] and in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals by University of Southern California, where the protocol was approved by the Institutional Animal Care and Use Committee (IACUC) (Permit Number: 12007 USC). Transgenic FlipTrap Gt(desm-citrine)ct122a/+ line was obtained from a previously described screen in the lab [23], Tg(kdrl:eGFP)s843 line [24] was provided by the Stainier lab, and Tg(ubiq:membrane-Cerulean-2a-H2B-tdTomato) line was generated by injecting a construct containing tol2 transposable elements flanking the ubiquitin promoter, coding sequence for membrane localized cerulean, a short sequence encoding the ribosome-skipping peptide of Thosea asigna virus (2a) followed by H2B-tdTomato. Upon crossing appropriate adult lines, the embryos obtained were raised in Egg Water (about 60 μg/ml of Instant Ocean and about 75 μg/ml of CaSO4 in Milli-Q water) at about 28.5° C. with addition of about 0.003% (w/v) 1-phenyl-2-thiourea (PTU) about 18 hpf to reduce pigment formation [28].
